体外循环手术前后病人血清sIL-2R水平和T淋巴细胞亚群、NK细胞的变化及临床意义

目的：通过检测心脏体外循环（ＣＰＢ）手术前后病人血清中可溶性白介素２－受体（ｓＩＬ－２Ｒ）、Ｔ细胞亚群、自然杀伤细胞（ＮＫ），观察分析心脏ＣＰＢ手术病人细胞免疫的影响及其临床意义。方法：选择２４例风心病择期换瓣病人，术前、ＣＰＢ１０分钟、ＣＰＢ２小时、术后第１天、３天、５天检测血清ｓＩＬ－２Ｒ水平、Ｔ细胞亚群及ＮＫ细胞活性。术后第１天血清ｓＩＬ－２Ｒ水平升高，ＣＯ４（辅助细胞）活性明显降低，ＣＤ４／ＣＤ８（辅助细胞／抑制细胞）比值下降，ＮＫ活性降低；并且ｓＩＬ－２Ｒ与ＣＤ４、ＮＫ活性呈负相关。结论：低温心脏ＣＰＢ手术对病人免疫机能有不良影响，临床应采取相应措施，改善术后病人的免疫机能。     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.     

活化性杀伤细胞免疫球蛋白样受体阳性的NK细胞对靶细胞的杀伤作用

Objective To study the killing effects of the killer cell immunoglobutin-like receptors (KIR) KIR2DS1-positive natural killer (NK) cells against acute, myeloid leukemia (AML) target cells, and to explore the KIR, human leucocyte antigen-Cw, and Cw loci (HLA-Cw) of NK cells, and, HLA-Cw of target cells mismatches destruction mechanism. Methods High-purified NK cells separated by DNYAL bead negative selection from healthy donor peripheral blood were taken as effector cells, and the freshly isolated bone marrow mononuclear cells from newly diagnosed AML patients as target cells. The anti-CD158a, CD158b monoclonal antibody was used to block inhibitory KLR receptors of NK cells (such as: KIR2DL1, KIR2DL2, KIR2DL3). The NK cells cytotoxicities against target cells before and after KIR blockades were detected by MTT reduction assay in vitro. Polymerase chain reaction and sequence specific primers (PCR-SSP) genotyping techniques were used to detect HLA-Cw, KIR gene of the healthy donors and patients. NK cells and target cells were divided into group C1 (expressing HLA-Cw 01, 03, 07, 08, 12, 14, 16 alleles), C2 group (expressing HLA-Cw 02, 04, 05, 06, 15, 17, 18 alleles) and the C1/C2 group (co-expressing the alleles in C1 group and C2 group) based on HLA-Cw. Results The purity of NK cells analyzed by flow cytometry was (90.8±6.08)%, and the killing effects of NK cells were significantly enhanced after inhibitory KIR blockades (t=-3.00, P=0.005). The cell lysis in C1 group, KIR2DS1+ NK cells against C2 group target cells (57.37±1.40)% was significantly higher than that against C1 group (44.19±4.67) % and C1/C2 group (36.77±6.56)% target cells (F= 11.87, P = 0.021), furthermore the differences in cell lysis among the above three groups become more evident after the inhibitory KIR blockades (F = 18. 72, P=0. 009). Conclusions The activity of NK cells was increased after the inhibitory KIR blockades, and the killing activities of KIR2DS1+ NK cells were higher than KIR2DS1 NK cells. The incompatibility between KIR, HLA-Cw of NK cells and HLA-Cw of target cells mediated NK alloactivation, realizing "missing self" recognition, and enabling NK cells cytotoxieity against targets.